HU230294B1 - Use of il-18 inhibitors for treating or preventing cns injuries - Google Patents

Description

The present invention relates to the field of pathological conditions of the brain. More particularly, the present invention relates to the use of an IL-1A inhibitor for the treatment and / or prevention of central nervous system injury, preferably salt traumatic brain injury.

In 1489, an endotoxin-induced safety deposit from mouse spleen cells was described. which activates imeroföron-γ (IFN-γ) (Hakamura et al., 1989). A. lymphadenoma does not directly induce IPN-γ, but rather does IL-2 or mitogenogenase as costimulants. Mouse eye phenol after endooxin treatment has attempted to purify the activity. and an apparently homogeneous 5O-S5 kDa protein was obtained. Other cltokines may act as costly to produce IFN-γ, but since IL-1s neutralizing antibodies to IL-4, IL-S, IL-oh or TNF were unable to neutralize serum activity , so different from these factors - in 1995, the same researchers reported that endotoxin-induced IFbl-Y production was also found in the liver extracts of preconditioned mice (Ekamura et al., 19: 5). in this model, the liver-derived macrophage population (RupSor cells) was proliferated, and in these mice, a small dose of bacterial lipopolyseal bar (LPS), which is non-lethal in mice with non-lethal IFN-′ -inducing (akunak). (dl '), subsequently called interleukin-iS. (IL-18) was purified to homogeneity by degenerate oligonucleotides from 1200 g of A / l treated mouse liver A purified lL-1S mimic acid sequence. IL-18 and interleukm-1.2 (IL-12) mRHSe are readily detectable in Kmpile cell activated activated macrophages of 11,18,157 aromatic acids, 18-19 kDa, which did not show obvious similarity with a single peptide in the databases. Recombinant 1L-18 induces IFN-γ more efficiently than 11-12, apparently by different pathways (Micallef et al., 1 '>%) Induced by ondoioxin serum ktb, to the lips it is

.. '?

judgment IS alone does not induce ÍFNA. but primarily kostinwhmsken with six mrtogens or IL-2. IL-U stimulates T ^ ej * -polyferado _} via a Π, -2-dependent pathway, and stimulates Tbl-ci toxin production in vitt-o, and IL-O-vC in combination produces a synergistic effect on IFN- γ for increased production .5 (Máitszewsö et al. 1990).

After cloning of the murine spleen, the hNman enNA sequence of IL-18 was reported in 1996. (Üshlo et al., 1996).

Cloning of judgment-18 from affected tissues and assaying for; IL-1§ gene expression revealed a close association between this eitokine and an autoimmune disease. In non-m / m diabetic mice (HOD), 'chrono-obese diabetic mice' spontaneously develop autoimmune insulins and diabeies, which can be accelerated and synchronized with a single mole of cyclo-star. reverse transcriptase PCR revealed the presence of IL-1S-niRNA in the pancreas of NÖO mice during early stages of insulosis. The IL-18 synth mBNS increased dkiöfoszfamíd utáa treatment _and preceded and IFM-y-mlOvS növeke15 of dose, and was basified with diabetes quickly. Interestingly, this is kinetics. It resembles the kinetics of IL-12 ~ p49 mRNA, which results in close correlation of individual ruRMS levels. Sequencing after cloning of IL-1b-eDNA from the pancreatic gland BNS showed identity to the IL-13 sequences cloned from Knpfier cells and carrier preactivated macrobeads. Macrophages from the HOD mouse also reacted with Π ..- 18 gene expressors

20-cyclophosphamide, whereas Bibib / c-ege treated similarly? this is not the case with the resulting macro branches. Thus, IL-18 expression is abnormally regulated in autoimmune NOD mice and is closely linked to the development of diabeies (Rothc et al., 1997).

B-I§ plays an important role in immune regulation or inflammation by enhancing the functional activity of Fas ligand on Thl25 cells (Centi et al., 1997). 1L-.18 is also expressed in the adrenal cortex, and thus may be a secreted nonimmune modulator, which plays an important role in the post-stress control of the immune system. 1986).

, ¾ wo is formed into the cleavage of Η, -18 pro-IL-1S and occurs as a lake due to its endogenous activity. in ecum IFN-γ production and I, PS-mediated. The mature 8L-I8 is formed from the precursor by the εL-I β converting enzyme kteem flt1 beis-converting enzyme, ICL, caspase ~ l>.

of two components, which act as a single binding site for binding to your knee. The strong and weak affinity binding sites of IL-18 have been identified in mouse

II, - 12 stimulated i-celloken (Yoshimoto et al., 1998), suggesting that it is a multi-chain receptor complex. To date, two lobe receptor units have been identified, both belonging to the IL-1 receptor family (Famei et al., 199b). Signal transduction of IL-18 also involves activation of '19 NF-κΒ (DiDonato et al., 1997),

Recently, soluble proteins with high affinity for IL-18 have been isolated from human urine and have been described with human and murine cDHSs (Növitek et al., 1999: WO 99A} 9963k A), a protein binding protein (IL-1B). , "IL-18 bbfewprotein").

IL-18BP is not the exiraclucleotide domain of one of the known L-18 reepeptors, nor is it a secreted, naturally occurring protein in the beverage. It belongs to a new family of secreted proteins, including: a. is a member of the ntogo many proteins encoded by Tóx virus with strong homologies to K18-8BP (Novick et al., 1999). IL-18BP is constitutively expressed in the spleen, belongs to the immunoglobulin superfamily, and has a limited homology to the IL-1 type II receptor. Its gene is located at the 11 lqI3 site of the hippocampus and no exon coding for the transcriptome domain has been found in an 8.3 kb genomic sequence (Novick et al., 1999),

The four human and two murine isoglobulins of Ik-ISBB, derived from mfeHS excision and found in various eDÁS libraries, were expressed. have been purified and evaluated for non-binding to their binding and biological activities of IL-18 (Kim et al., 2000). A. bamánemdefe 11, -IBBFa-taste gene (IL-18BPa) showed the highest affinity for IL-18 at a fast on-off rate ("OB-mte")> a slow off-rate (offirate) and a 399 pM docking constant (K (d) l IL-I in SöPc and IL-18BPa has the same Ig domain except 29 C-terminal amino acids, but *

4lL-18BPe has a K tdj of only one tenth (2.94 nM). IL-1SBPa and 1L-18BPc, however, do not overfold the IL-18 by> 95% bar in a double molar excess. IL-18BPb ··· and ILL18BPd isoforms lack a complete Ig domain and the ability to bind IL18: -boz or neuralysis. Mouse-derived IL-1SBPc and IL-1Sf1Pd-expressing mice, which have the same Ig-doxnene, also centralize> 95% in mouse-derived IL-1S in a double molar excess. However, mouse-derived H18-8BPd, which contains a brown C-lobe sample shared with brown IL-1 & Blife, also does not nababolate human IL-1S. Large, mixed, electrostatic and more hydrophobic binding sites have been identified in the IL-18BP Ig domain, which may explain the strong affinity binding to the ligand (Kim et al. 2000).

Intra-brain injuries (IBI, traumatie bram mjury '% or simply head injuries, or closed head injuries (Cili. Closcd head tendon)) refer to injuries to the central nervous system where brain injuries are caused by external impact to the head. Mostly car or bicycle accidents but can also be the result of suffocation, heart attack, stroke and infection. This type of traumatic brain injury is due to a lack of oxygen or blood supply to the brain and is therefore referred to as "noxic injury".

Closed head injury occurs when you hit the head as an example! in the event of a motorcycle accident or fall. In this case, the skull collides with a stationary object and the brain in the collarbone rotates or twists around its axis (brainstem), causing local or distant damage. And the brain - surrounded by a liquid and:

soft material floating in it - it could hit the skull which could cause further damage.

Immediately after the trauma, there may be a period of unconsciousness that may last for minutes, weeks, or months. Traumatic Injury: A patient's brain may suffer from multiple parts of the brain: injuries or bruises. This is called diffuse injury or non-projectile damage to the brain. The types of agvt lesions that occur during non-projectile casualties may be primary \ eg \ secondary.

> # * ** «« · **

Primary brain damage occurs at the time of injury, e> -oMm at the site of impact, especially if a skull fracture has occurred. major lesions may be associated with mtraeerebral haemorrhagic lesions or cortical injury, DrSúz axonal injuries are the result of neural processes caused by the rotating movement of the matter within the skull: shear or stretching of the spine, These may be small, haemorrhagic lesions or micro

Secondary brain damage can be the result of complications occurring after the time of injury, such as intracranial haemorrhage, traumatic damage to the extra-cerebral arteries, intra-eranic hernia, hypoxic brain damage, or meningeal tis,

A. Open Head Injury ”a. visible injury to the head, and may result from a gunshot wound, an accident, or an object entering the brain through the skull (brain projectile injury '). This type of head injury is very likely to damage a specific area of the brain, $ The so-called medium brain injury does not cause unconsciousness and only short

It may cause temporary dizziness or confusion. Persons with non-coma-related brain injuries, although requiring only minimal medical attention, may experience symptoms similar to those of survivors of coma injury.

In response to trauma, changes occur in the brain that require monitoring to prevent further damage. After a serious head injury, the size of the brain often increases. This is called cerebral swelling and occurs when the amount of blood in the brain increases. During the course of the disease, water builds up in the brain, which is called cerebral edema. Both cerebral swelling and cerebral edema result in overpressure of the brain, which is referred to as intra-arterial pressure (ICP).

Coma is a prolonged period of unconsciousness that immediately follows a traumatic head injury.

-ό4 «*» «9 9

Φ 4 * 944 »X * * .Χ · 4

9φ »9 * $ * * * Λ« fc * <* φ * φφ- # ·

Coma has different levels. Coma levels can be measured by improving the responsiveness of the person with the head injury. In the acute phase of head injury, the Gl®> ~ gow Scale (Glasgow Coma Scalefo) can be used. As the patient's condition improves or stabilizes, the ^fS Raneho Los Amigos scale, which measures levels of cognitive thinking (understanding and reasoning), can be used.

Brain injuries often result in persistent debi Ina, such as post-traumatic epilepsy, persistent vegetative state or post-traumatic dementia.

Spinal cord injuries are another type of central nervous system injury (ÖMS injury, 'foentral: nervous sysfem'). Most patients admitted to hospital for bilateral or complete paralysis are the cause of spinal cord injury. More than 8Ö ° / e ~ our Road? the result of an accident. Clhukically, two major groups of injuries are recognized; open injuries and closed injuries,

Open injuries cause direct trauma to the spinal cord and nerve roots. Injuries to the lawsuit can cause edema and hemorrhage. Most spinal cord injuries are closed injury and are usually related to fracture or protrusion of the spinal column, which is usually radiologically measurable kk. Spinal cord injury depends on the extent of bone injury and has two main conditions: primary damage, which secondary damage, which is esoteric hematoma, infarction, infection and edema.

Post-effects of spinal cord injuries; ascending and descending nerve fibers, asterogradic degeneration, systemic effects of post-traumatic stringomyah and bilateral paralysis, such as urinary and chest infections, pressure on pain; and isotonic atrophy,

A. The pathology of french brain injury is very complex, and still little known. Experiments in the last physical year have shown that systemically and locally released eutokines play a role in brain injury after brain injury and have been shown to have dual effects on prominent human cytokines such as TNF, IL-6, or 11-8. based on time-dependent, volatile and adverse effects of mediators (Morganti-Kossmann et al., 1997: Kossmann et al., 1997; Shohstni

V * Λ · * * * $ * * κ »* $ # · * $% 'k * ♦

V * ·. # ** · ** · * et al., 1999; Seherbel et al., 1999; Whalen et al., 2009). As. the. Previously, we have disclosed that a recently recognized member of the IL-1 eletin family is IL-18. Recent studies have shown that 1L-1K is a carrier in the central nervous system of mice, rats and human subjects (Culhane et al., 1998; Jander and Stoll, 1998; Fsanz et al., 1999; Fassbender et al., 1999; Wheeler et al., 1999). et al., 2009), and is constitutively expressed in the primary culture of astroelets and mycoglia but not in the culture of neurons (Centi et al., 1999); Elevated levels of 1L-1S were not observed in CSF of 19 patients with multiple sclerosis (MS) (Fassbender et al., 1999). .MS patients generally had low levels of intrafoecal IL-18, while a higher level of IL-18 mKlbfi was observed in experimental Lewis rat animal model of autoimmune eneephalomyelitis (BAE). and SfoH, 1998). The expression and functional importance of IL-1S15 in neuroarray has not yet been investigated,

The invention will now be briefly summarized. The present invention relates to its pathophysiological role in 11 to 18 CNS diseases. Based on the finding that treatment of mice with IL-18 inhibitors for up to one hour or three days, experimental closed head injury (CH1) improves healing (1) and reduces brain damage compared to control animals. The present invention relates to the use of IL-18 littermates for the preparation of a medicament for the treatment and / or prevention of central nervous system injury, preferably traumatic brain injury.

Combinations of the IL-18 inhibitor with interferon and / or TNF and / or anti-inflammatory and / or antioxidants are also contemplated by the present invention. A gene therapy approach for the delivery of IL-1S-incubation to diseased tissues or cells is contemplated, and the invention further provides the use of nL-18 encoding sequences for the treatment and / or survival of a CKS lesion. , genetically engineered * <<

* «* R» · use of tort cells: to prevent and / or treat CNS injury to express IL-18 inhibitors in cells.

The figures are briefly described below.

Figure 1 depicts a brtogram of mtracetebral IL48 (ng / ml) serum NMR in whole brain (striped), fish hemisphere (black) or right hemisphere (gray) under various conditions.

THE 2. Fig. 4A shows the development of NSS (Neurologica! Severity Seore's Neurology Severity Score) at 1 hour (b), 24 hours, 72 hours or 168 hours after trauma follow-up, or 50 gg, 1 hour post-injury. , given IL-ISBF (squares) or injection of vehicle only (control, circle),

Figure 3 shows the ANSS values at 24 hours, 72 hours, or stroke hours after the stoma, or 50 pg, 1 hour after the injury, i.p. given IL-1 & SF (squares) or injection of vehicle alone (control, circle). Figure 4 shows the ANSS values at 1 hour, 24 hours, 3 days (d), 7 days, or 14 days measured after or 50 pg IL-1 SBF bp. given either in a single dose (rhombus) 3 days after the trauma, or in two doses 1 hour and 3 days after the trauma (square), or by injection only with the vehicle (control, triangle),

The invention will now be described in detail. The present invention is based on the discovery that the IL-IS inhibitor has a statistically significant beneficial effect on the recovery of brain injury in a mouse model of closed head injury. In the context of the invention, it. also found that after 11, -18 teath brain lesions, it is regulated at a higher level in the brain and in the cerebrospinal fluid, suggesting that this proinibunma toric cifokine plays an important role in the ontogenesis of brain injury,

The present invention thus relates to the use of IL-18 inbifene in the manufacture of a medicament for the treatment and / or prevention of central nervous system (CMS) injury.

«» ♦ *> *

The present invention also provides the use of an IL-18 inhibitor for the manufacture of a medicament for the treatment and / or prevention of complications and after-effects of CNS injury.

In preferred embodiments of the invention, the CNS injury is a transgenic brain injury or a closed lumbar injury.

In other preferred embodiments, CNS walking is a spinal cord injury,

In a further preferred embodiment of the invention, the brain injury is of vascular origin.

As used herein, the term "central nervous system" refers to any injury to the brain or spinal cord, irrespective of the age of the injury and the underlying cause. Causes include mechanical baiting or infection. The OES lesions, their clinical symptoms and consequences are described in detail above. For example, CMS-Injury a. trauma, or any other damage to the brain or spinal cord, and can be referred to as non-neuroma.

Brain injuries can be, for example, the following, or one, two, or more of the following; 1. wrinkle destruction. 2. impairment of cognitive skills, 3. use of language ,. 4, memory impairment, 5, lifestyle disorders, he. motor disorders, 6, any other neurological disorder,

For example, spinal cord injury can result in bilateral or complete paralysis.

Complications or after effects of CNS injury can be treated and / or prevented according to the invention. The complications and after-effects of brain injury have been described in detail above. Examples include, but are not limited to, coma, meulugitis, post-traumatic epilepsy, post-traumatic demaphy, nerve fiber degeneration, or post-traumatic lymphoma or haernorrhagia.

The present invention also relates to the use of 11,18-tnb libraries for the preparation of a medicament for the treatment and / or prevention of any vascular brain injury such as hypoxic brain injury with cerebral infarction, isehaemla, eerebrovaseulsris.

-neither " * *

The tea .a treatment and prevention terms CNS damage one or more symptoms or _ofebiak,: partial or complete, preventing, inhibiting, alleviating, improving or visszaferditása accompanying € NS ~ Injury symptoms, diseases or complications expertise in Lasznábufc CNS injury has already handling the substances according to the invention a

It is given after the onset of the disease, and, in the case of prevention, the administration of substances before the symptoms of the disease can be prescribed to the patient.

The treatment of CNS injury according to the present invention is preferably carried out. Preferably, for the treatment of CMS injury, the IL-15S inbibdor after the CNS injury can be administered as soon as possible, for example in the first hour after the injury. However, in the Examples section below, it will be shown that one of the 11,108 inhibitors has a beneficial effect on brain injury, even if given three days after the brain injury, so that the inhibitor of IL-18 ~ is preferably used to treat CNS injury. three days after injury.

According to the present invention, the term Π, -18-hihitor is understood to mean a molecule that interferes with the production and / or activity of IL-18 by reducing, reducing, or partially, substantially or completely inhibiting the production and / or activity of IL-18. or inhibit it. By IL-10 inhibitor is meant inhibitors of IL-18 production and IL-18 activity,

Inhibitors of production can be any molecule that adversely affects the synthesis, processing, or maturation of IL-18. Inhibitors of the invention include, for example, mucil-γ-bbs gene expressions that reduce or prevent . Π ..1 results in transcription of mRNA or degradation of mRNA, whites that do not: cause proper cleavage, or partially or substantially inhibit the secretion of 0, -18, lL-18 by pro-enzymes that immediately degrade symmetry, IL-18 cleaving proteases that produce mature IL-18, such as easpase I Inhibitors, and the like.

For example, an inhibitor of IL-18 activity may be an IL-18 antagonist. The antagonists either bind to the IL-1d molecule itself or are sequestered with sufficient activity and specificity to partially or substantially neutralize IL-18 or oxII-18 «V *«

X χ. «Χ * Ο * χ *» * ♦ * χ «χ * ♦:

* ♦♦ * ♦ * «« «· **« «« felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős felelős♦ ♦ ♦ IL ♦ felelős when ILII 8 binds to its receptor.

Inhibitors of IL-18 activity may be soluble β-18 receptors or molecules that mimic reeeptoro5, or inhibitors of IL-18 receptor or anti-1L-18 antibodies, such as monoclonal or polyclonal antibodies, or any other a molecule that prevents the binding of IL-18 to its target, thereby reducing or preventing the secretion of IL-18 mediated intra- or extracellular reactions.

In a preferred embodiment according to the method of the IL-18 inhibitor can be selected ;: inhibitors of caspase-I _(1CE): antibodies to 1L-18, either an antibody against IL-18 receptor subunits, inhibitors of the IL-18 szígnálúivonalának, IL -18anfegonists that are competent or bind to IL-18 and inhibit IL-1Sreeeptor and IL-18: -binding proteins, isoforms, artificial metals. fused feces, functional derivatives, active fractions or circularly permutated derivatives or salts thereof.

As used herein, the term "LL-18 ~ fusion proteins" is used as a synonym for the term "L-1888F ^ts, " and is used herein to refer to IL-1S binding proteins as described in WO 99 / 09-63 or Novick et al., Noviek et al. 1999}, including splicing and / or isotherms of IL-18 lobe proteins as determined by Kim et al. (Kim et al., 2000). Preferably, 1L-18SF a and c according to the invention A. The proteins useful in the present invention may be glycosylated or non-gh wells, may be derived from naturally occurring females such as urine, or may be produced recombinantly, may be used in recombinant expression systems,

E. G. E. ooAhan, or eukaryote, and preferably in mammalian expression systems.

A preferred cell line for the III, 18 inhibitors of the present invention is Chinese Hamster Ovary cells (CHO cells, Thinese hamster ovary).

»· 44 * * Κ» * * · »* fc 4 fc · 4 4 *« fi »94 4 fi» »44

444 4 Μ X ♦ * fc · »: ** 44 fc *«

Recombinant production of IL-18 inhibitory, when expressed recombinantly in a mammalian cell or mammalian cell line, is preferably performed in a non-streaky cell culture medium.

As used herein, the term "mOW" refers to analogs of IL-18BP or watermelic 1SBP in which one or more amino acids of naturally occurring IL-18BP or viral-derived IL-1SBP are substituted or deleted with different amino acids. more amino acids are added to the natural sequence of E. 18BP or 1LI8BP of water origin without significantly reducing the activity of the resulting products in the wild type. IL-1 & SP or viral IL-18BP ~. These maternities may be prepared by known synthesis technologies and / or site specific mutagenesis technology or any other known suitable technology.

At each of these levels, its amino acid sequence is a sufficient copy of the IL-18BP or viral-derived IL-18BP winoic acid sequence to have substantially the same activity as IL-18BP. One activity of I L-.18BP, tip hegy ..-1 S. When the malay has essentially IL-18 binding activity, it can be used for purification of IL-18, for example, by means of aromatic chromatography and thus considered to have substantially the same activity as IL-18BP. It's that. whether a given m-cycle has substantially the same activity as IL-18BP can be determined by rally experimentation, e.g., a simple sandwich competition assay with nmtein can be used to determine whether or not it is appropriately labeled with IL-18, such as a radiolabel or Elise vízsgábt. Single, functional studies to evaluate the biological activity of IL-18BP a. WO 99/09063 is described, for example, in Example 2 (binding to IL-18 is evaluated by cross-linking) or in Example 5 (inhibition of IL-18 induced by tNF-gamma in mono-chemical versions).

In a preferred embodiment, at least 40% of each of these lymphomas is identical or homologous to either the homologous sequence of IL-1BB or the virally encoded IL-18BF. More preferably, at least S0%, at least 60%, at least 70%, at least ί

-.13 ♦ ♦: * ♦ ♦

». * & Λ t» * * Κ

Wt% or most preferably at least 90% identical or homologous to the homologous sequence of either IL-ISBF or virus encoded 1L-ISBP

Suitable IL-1 SEP muteins or virus-derived IL-1hB1? Mules or their encoding nucleic acids for use in the present invention include, for example, finite numbers of substantially 5 leo sequences, such as substitution peptides or polynucleotides, which can be routinely prepared by one based on the teaching and guidance of the description ..

Examples of mnteins according to the invention are nnkleic acid. DNA or RNA encoded proteins that hybridize to nucleic acid with DHS or RNA encoding the inventive IL-8 inhibitor in moderately or severely lysing conditions. The term stringent conditions refers to those hybridization and subsequent washing conditions, which are commonly termed stringings by one skilled in the art. See, for example, Ausubel et al., Current Frooeols in Molcettelar Biology, p. supra, Intsrseience, Nt §6.3 fc §6.4 (19 & 7 _;. 1992) and Samferook et al., Sambrook, 1 C Fritseh, E, p. and

Mamaris, T ,, Molecnlar Gtening: A. Lahoratory Manual, Gold Spring Anywhere Lahoratory Press, Coki Sprirtg Earber, NY (1989)].

Stringent conditions include, but are not limited to, washing conditions having a temperature of 12-20 ° C lower than the calculated Tm of the hybrid under test, e.g. With 2 x SSG and 0.5¾ SDS for 5 min, 2 x SSG and 0.1% SDS20 for 15 min with 9.1% SSG and 0.5% SDS Sv ^ C- wash for 30-60 minutes followed by 0.1x SSG and 6.5% SDS at? ⁹ G for 30 minutes :. One skilled in the art will recognize that stringent conditions will depend on the length and length of the DNA sequences, ohgonkleotide probes (e.g., 10-46 bases) or mixed oligonucleotide probes. If probes are used, tertiary methylmmonium25 chlorodifc (TMAC) is used for flooding. Instead of SSC. See Ausubel (stgw).

Preferably, in one embodiment, each of these lineages is at least 40% identical or homologous to SEQ ID NO: 1, 2, or 3 in the attached SEQ ID NO: 1; nicely at least SÖ% ~ han, at least 06%, at least 70%, airX X Φ ♦ * ♦ * • 'ϊ «»

-14 <80% or more preferably at least 90% identical or homologous to SEQ ID NO: 1, '2 or 3

Nuclear Identity: Refers to the relationship between two or more polypeptide sequences or two or more polynucleotide sequences, which is effected by alignment of the sequences. Generally, identity is a. based on the exact match of the n-nucleotide-nijkleodd or the amino acid-ananoic acid of the two potty nucleotide sequences or the two pepper peptide sequences.

For those sequences where there is no exact match,% identity can be determined. In general, the two sequences to be compared are aligned adjacent to each other so as to obtain maximum alignment between the sequences, whereby holes can be inserted into one or the quality sequence to enhance the degree of alignment. The% identity can be determined over the entire length of each of the sequences being compared (so-called cohort alignment), which can be used primarily for sequences of the same or very similar lengths, or more rigorous determinant lengths (so-called local alignments). applicable.

Methods for comparing the identity or homophagy of two or more sequences are well known to those skilled in the art. For example, the programs included in Kfeconsm> & </ ausee AnafysA Pncixe Version 9.1 (Devereux, J, and míssL 1934), e.g.

BESTFTT and O'.AP programs are applicable to two. polynucleotide percent identity and percent polypeptide sequences percent identity and percent decomposition, BESTFIT Srníth and Wutennan (1.981) '' use a local homology algorithm, and looks for the most similar single region between the two sequences. Determine the identity between sequences, and / or other similar programs are known in the art such as BLAST szeritfo progtárncsalád (Áfeschuk S, F. et al 9ik yl ° l S i ts et ^l'u í '\ IUI website vtic-ztui cdvfo el wmv.ncbknte.níb, gov), and the FASTA-pmgram (Beamon, W. Fe, 199Cf; Pearson, 1983).

** * χ χ * »*« »* ·« * «.

ί *? .... *,; · * »

Advantageous changes for the inventors of the invention are the so-called conservative substitutions, the conservative amino acid isfunctions of the IL-1SBP-polypeptides or proteins, or the viral-derived 1L18BPs, which are synonymous with the following properties:

substitution between the two molecules preserves the biological hmkdo of the molecule (Grantbaur, 1974). It will be appreciated that amino acids may be inserted and deleted in the above sequences without altering their function, preferably if the dimers or déctions affect only a few amino acids, e.g. less than thirty and preferably less than ten, a functional conformation such as cysteine. Hunting proteins and surgery for such deletions and / or tendons are within the scope of the present invention.

Preferably, the synonymous amino groups are those in Table 1. More preferably, the synonymous amino groups are those defined in Table A, and most preferably amino groups.

the synonymous amino groups in Table 3 are confused

amino acids

Ser Arg Len Pro Thr

Fall on Val Gly

Phe

Tyr

Preferred donors of X-fever

Synonym group

Ser, Tbr, Gly, Asn

Arg, Gin, Lys, Glm His be, Phe, Tyr, Met, Val, Len

Gly, Ala. Tbr, Pro

Pro, Ser, Alá, Gly, Hís, Gin, Thr

Gly, Tbr, Pro, Alá

Met, Tyr, Phe, lie, Leu, Val

Ab, Túr, Pro, Ser, Gly

Met, Tyr, Phe, Val, Len, He

Trp, Met, Tyr, Here, Val, Len, Phe

Trp, Met, Phe, lie, Val, Lem Tvr «» ♦ * »**

S * *

• 16-

L, ys His Gin Asn bcr, í hi \ Lys Gin, Lys, Gin, Thr, Arg, His Gin, Lys, Asn, His, TI Gin, Asp, Ser, Asn Irish, Arg, Gin Lys Gin, G In, His, Arg, Ly • -'8 asp Gin, Asn, Asp Gin Asp, Lys, Asn, Gin, H A, Arg. Gin Met Phe, De, Val, Len, Me t Trp W

Table 2

Preferred group of synonymous amino acids?

Amin osav 57n? orn.n? Group? Arg His, Lys, Arg · Mf. Pro Thr AaV-'j ί iíA> _x iyLVt Ab, Pro Thr ab With Pro, Ala Val, Me ?, Se Gly Gly Neither It's McL Phe, Val, Phe Met, Tyr, Be, Len, 1AT Phe, Tyr Gys Ps. Ser PbS Gin His, Gin, Arg Gin, Gin, His Asn Lys Asp, Asn Lys, Arg THE Gin Asp, Asn Gin, Gin Me? Met, Phe, Se, Val, IRP Trp

Leu

Phe

Len xt ** »> ·· '' s ·· * · & * 9

-r

amino acid

Ser

Price Latvia Pm Tbr Ab

Val Ik Ikhe Table

More favorable groups of synonymous synonyms Synonym group Ser

Len, lle, Me

Pro

Thr

Ah

Gly

Ik, Met, Lett Phe

Cys: Hrs Gin

Asa

Lys

Asp Glu Met Trp

Cys, Ser

IF

Gin

ASN

Lys

Gin

Met. Ile, Len

W

White: Examples for the suppression of amino acid standard transitions, such as those for the production of artificial IL-88P polypeptides or proteins or vhusercdetu IL-1BBP-rmkem, which are known in the art, e.g. U.S. Patent Nos. 4,588,585 and 4,737,462 to Mark et al., 5,116,943 to Koths et al., 4,965,195 to Namen et al., 4,879.11 to Ghong et al. .691 (Lee et al.). No.: in U.S. Patents and Lziunel ♦ ** * »<* * ♦ ♦ ♦ *» »» «« * ** ** ♦. # ♦. * »

-18Shase proteins disclosed in U.S. Pat. No. 4,94,584 (Shawés et al.).

The term "fusion protein *" refers to a polypeptide that contains IL-18BP-1 or viral IL-ISBP, or a synthetic metal or fragments thereof, fused to another protein, e.g. stays in your body fluids for longer. Thus, IL-18BP or viral-like IL-18BP can be fused to another protein, a peptide peptide or the like, such as Irumunglefeuhn or a fragment thereof.

According to the disclosure, the term "functional bar derivatives" refers to derivatives of IL-18BP or viral IL-18BP and their rauteins and fusion proteins which are N- or

Functional groups on C-terminal groups or side chains of amylic acids can be prepared by prior art devices and are within the scope of the invention as long as they retain their therapeutic utility, i.e., they degrade the activity of a protein substantially identical to that of II-18BP or viral IL-18BP. , and do not cause toxic properties to preparations containing them.

These derivatives may contain, for example, polyleneglycol glycol side chains which may mask antigenic sites and extend the residence time of IL-18BP or virus-derived? the aldehydes of the free amino groups of the amino acids formed by the reaction of the amides with the acyl moieties of the free amino groups of the amino acids with acyl moieties (e.g., alkoxyl or carbocyclic atoll groups) or free hydroxy groups (e.g., hydroxy groups of serine or threo groups).

The 1L-18RP or viral .1L-ISB műiéinek ?, and fused active proteins írakcióiként ^'1 of the present invention fehérjeniolekula pohpepödláncának any feagmense or precursor therefor is alone or together with associated molecules or amino acids, e.g., sugar or phosphate groups, or fehértemolekula or a self-aggregate of groups of sugars, provided that. the reaction has essentially the same activity as. 11, -1 SBP-vst

-19 '* ·> ♦ «♦ V ♦ X;» * «X *: *: ♦ ♦ *> χ ♦» * ♦ * «4 * * **

As used herein, the term "salts" is understood to mean both salts of the carboxylic groups of one hundred lL-18 inhibitory molecules and the acid-addition salts of amino groups or analogs thereof. Salts of carboxyl groups may be formed by methods known in the art and include, for example, inorganic salts such as sodium, calcium, ammonium, iron, and omits and the like, and salts with organic bases such as antines such as trialanolamine, arginine or lysine, piperidine. , salts with procaine and the like. Acid-addition salts. salts with mineral acids such as hydrochloric acid or sulfuric acid; and salts with organic acids such as acetic acid or oxalic acid. Of course, all salts must retain the biological activity of the ÖPN relevant to the invention. that is, stimulate the proliferation of oligodendrodes,

In another preferred embodiment of the invention, the inhibitor of 11, 18 is an antibody against 11, 18. The IL-18 antibody may be polyclonal or monoclonal, lamellar, humanized, or entirely human. The recombinant antibodies and their phage fragments exhibit high affinity and small toxicity in mm II, -18 winters. The antibodies useful in the present invention are characterized in that treating patients for a sufficient period of time will relieve a good amount of urine or any symptom or group of symptoms associated with the pathogenic condition and are slightly toxic.

Neurizing antibodies are in animals, such as saliva, goat or mouse

2E can easily be produced by immunization with 11, -18. Immunized mice are cytopathic sources for the production of B cells for plaque production, which can then be cultured to produce large amounts of monoclonal antibodies to 1L-18.

Iamera antibodies are like that. immunoglobulin molecules which are characterized by two or more segments or portions of different animal species. Generally, the variable region of exhaustive antibodies is derived from non-human mammalian antibody, such as a murine moclonal antibody, and the constant region of an immunoglobulin is derived from a human tmmnnglobuHn molecule. Preferably both sections and the combo * «Φ ♦ * * φ« a φ χ, χ * * W «» φ * «* ♦ ** φ> φ ****« Φ <φ Ν «· * ** * · * * Χ Λ · * «

A minor immuno-gene. as is easily determined (Ellíott et al., I994k Humanized antibodies are genetically engineered immunoglobulin molecules in which murine constant sections are human-identical, and murine antigen-binding sections are human and human-derived). in human organisms (Knigki et al., 1993).

In a further preferred embodiment, the IL-15S promoter material is a humanized 11,18-moiety. Preferred examples of anti-humanized IL-18 antibodies are described, for example, in European Patent Application Serial No. 09-04,600.

Another embodiment of douyos. according to IL-18, the linseed is entirely of human origin. For a detailed description of technologies for the production of human antibodies, see, for example, WO99 / S3649, International Patent Publication No. WO99 / S3649; Fully human antibodies are preferably recombinant antibodies produced in transgenic animals, such as xenoegers, which contain all or only parts of the entire functional human tag.

In another preferred embodiment of the invention, IL-1S-inhlhtlor 1L-18BE or isotherm thereof is mutant. fecundated protein, fenceionic derivative, active fraction or circularly permutal derivative, Are these isofen? surgery, fused proteins or fancdonal derivatives, mimic the binding to the biological active site of IL-18BP, preferably digest IL-18, and preferably at least substantially similar to IL-18BP. ideally the biological activity of such proteins is higher than that of unmodified 1L-18BF - Oh. Preferred active fractions have better activity than. They have IL-ISBPs or other advantages, such as improved stability, lower feinting or mimicry, or ease: they can be produced in larger amounts or easier to tile.

Variants / isoforms of IL-18BP and IL-splices) are available from WO99 / 99003 or available from Novick et al. (Novick et al. 1999) and Kim et al. .

Functional derivatives of IL-ISBF may be linked to a polymer to enhance the protein properties, such as stability, life span, bioavailability, and tolerance of the human organism. or its immunogenicity. To accomplish this, the functional derivative may comprise at least one moiety linked to one or more phenylethyl groups on one or more side chains. Examples of such functional groups are polyethylene glycol (PEG). The formation of FEG-Derivatives can be carried out by known methods, for example those described in WO92 / 13995.

Thus, according to a preferred embodiment of the invention, the FEG derivative of IL-1FBs and preferably IL-1SBP is formed,

The invention is a further embodiment; In a preferred embodiment, the 1E-18 inhibitor is a fusion protein comprising all or only part of the IL-18 binding protein and fused to a complete inmoglobulin or to an immobilis moiety. the fusion protein formed is IL-ISBF biological. it retains its activity, preferably binding to fL-18. The fusion may be directly either via a short linker (via a peptide linker, which may be short 1-3 amino acids long, or longer, e.g. 1.3 amino acids). The linker molecule may be, for example: the SEM (Glo-Phe-Mef) tripeptide sequence, or a 13 amino acid linker sequence comprising the .glu-Fhe-Gly-Ala-Gly-Iyeu-Val-Leu-Gly-Gly-Gfe-Fhe-Met sequence between this IL-18BP sequence and the immunoglobulin sequence, The resulting fusion protein has better properties, for example, it has a longer staying time in the teak decadishes. It has better gaxiriku activity, higher expression levels, or easier fusion protein purification.

In a preferred embodiment, E-18BP is fused to the constant region of the Ig molecule. Preferably, heavy chain sections, such as. human IgGi € B2- and “0% * 9Χ« «« ο «* V * X '♦ * * * ♦ ♦ * ♦» * »* 99 * * Χ * X« χ # $ _κ * 99

Fused to its CH3 ~ domains, Is-ISIM and immunoglobulin; is described in Example 11 of International Publication No. WO99 / 099003. Other isotherms of Ig molecules are also useful in the preparation of the fusion proteins of the invention, such as. fgO2 or IgCo isotherms or other Ig5 classes such as. IgM or IgA, for example. The fusion proteins may be monomers or multimers, hetem or Hornom ultimers.

Interferons are primarily known to inhibit viral replication and cell proliferation Iteration by the example of mterferoh-y! plays an important role in immune cataracts and inflammatory processes. Innerforon-β (ΙΡΝ-β, ¹-histone Interferon is believed to have hormone-inhibiting activity).

Accordingly, the present invention also provides the use of a combination of an IL-18 inhibitor and an interferon in the manufacture of a medicament for the treatment of CNS injury.

Interferons can also be linked to polymers to improve protein stability. For example, the conjugate between Interphon-β and Hell, Pollenethylene Ghkoi (PEG) is

International Patent Publication No. WO99 / 55377.

In a further preferred embodiment of the invention, the interferon is shotgun-β (LFN-β) and more preferably IFN-β 1a.

The production and / or action of Ik-18 is inhibited by. at the same time as imerferos, separately or separately.

Huoörfoxis has been reported in the literature to have both protective and toxic effects in brain injury (Shohami et al., 1999). Example 1 below shows that Injection of TNF into mice following severe brain injury caused a significant decrease in levels of 11, 18, which is shown. that 1SF may have a beneficial effect on the recovery from traumatic brain injury. In a preferred embodiment, the invention relates to the use of a JL-1S inhibitor in combination with INF, simultaneously, sequentially or separately, for the preparation of a medicament for the treatment and / or prevention of brain injury.

The combination of IL-5S inhibitors with JNF-α is preferred according to the invention.

Λ * * »·»:

* «Μ * ί '/ χ ♦ V * ¥ ♦ # *? ♦ *« «♦ <« * χ $: Φ χ- ♦ χ χ * Α * ** »χ, η» »« «

In a further preferred embodiment of the invention, the medicament further comprises an anti-inflammatory agent, such as NSIID (a non-steroidal anti-inflammatory drug). In a preferred embodiment, a COX inhibitor, more preferably a C0X2 inhibitor, is used in combination with the 1L-185 inhibitor. COX tendon sticks are known in the art. For example, specific COX2-inhibitors are disclosed in WO0 / 1/00229. The active components may be administered simultaneously, sequentially or separately.

Oxydial stress, preferably reactive oxygen species (ROS), also plays a role in the pathophysiology of brain damage (Sbohamí et al., 1997).

In a preferred embodiment of the invention, the medicament further comprises an antioxidant, administered simultaneously, sequentially or separately. Many antioxidants are known in the art, such as A, E, or E vitamins, or 5-amino acid or superoxide dismutase.

In a further preferred embodiment of the invention, the IL-1S inhihitori is from about 0.001 to about 100 mg / kg. or about 0.01-10 mg / kg body weight or about 0.1-3 mg / kg body weight or about 1-2 mg / kg body weight>

In another embodiment of the invention, IL-10H inhibits about 10 in the range of 0.1 to 1000 pg / kg body weight or 1 to 100 pg / kg body weight or 1 to 100 ng / kg body weight or 10 to 50 ug / kg body weight. we,

The invention further relates to the use of a nucleic acid molecule comprising a sequence encoding a JL-1 S inhibitor, a synthetic moiety, a functional derivative or an active reaction thereof, for the preparation of a medicament for the prevention and / or treatment of saCNS injury. for administration of the IL-18 inhibitor of the present invention when gene therapy is used. Preferably, the nucleic acid molectile is administered in a syrup.

ϊ »* 4»? * <* ** ^Χ . * * * · 24 · ♦ η

For example, in order to treat and / or prevent CNS injury, a gene therapy vector comprising an inhibitor of IL-18 production and / or effect may be injected directly into the diseased tissue, such as problems with systemic administration of gene therapy vectors such as vector dilution, target cells. or tissue, and side effects can be avoided.

The present invention also provides for the reduction and / or stimulation of endogenous production of IL-d8-inhibitory.r in cells in which IL-18mblbhor is normally not expressed or inadequately expressing the inhibitor. treatment and / or prevention. The vector may contain regulatory elements that function in the cells in which the inhibitor or IL-18 is to be expressed, such regulatory sequences or elements being promoters or promoters. The regulatory elements can then be inserted into the appropriate site of the genome by homologous recombination, whereby the regulatory sequence is operably linked to a gene whose expression is induced or stimulated in coffee15. The technology is commonly referred to as endogenous gene activation (LBndögenous Gene Achvatéon et al. International Patent Publication No. WO91 / W955.

It will be apparent to one skilled in the art that using the same technology, 1L-18 express kopmek megakadaiyo, h'ZÓ, II-18 uíhíbuo? alcove, frame uéiku. For the sake of affinity, a negative regulatory element, such as a silencing element, may be introduced into the glial locus of IL-18, which is; It results in down-regulation or inhibition of IL-18 expression. One skilled in the art will recognize that lowering or downregulating the IL-18 expressing agent has the same effect as traveling on the subtype of IL-18 to prevent and / or treat disease.

The invention further relates to a cell genetically modified in a manner that

IL-18 ~ inhibitor, for use in the manufacture of a medicament for the treatment and / or prevention of CNS injury.

-25 * *. ♦ · fc * ^{ -s <

The present invention also relates to pharmaceutical compositions, preferably pharmaceutical compositions for the prevention and treatment of inflammatory UNS injury, comprising a therapeutically effective amount of IL-5 and / or a therapeutically effective amount of interferon and / or a pharmaceutically effective amount of INF and / or containing an effective amount of a medicament and / or a therapeutically effective amount of an antfexidant,

In the composition, the inhibitor of IL-18-inbibifer easpase-1, an antibody against 1L-1S, is.

11, - Antibody against any of the reeeptoral units of Ig, a substance that inhibits the IL-1? -Gene pathway, an IL-18 antagonist that inhibits IL-13? as well as their isotherms, muteins, fused proteins, functional derivatives, active fractions or circularly over-derivatized derivatives having the same activity.

The above-described 1L-18BP and its isotherms, inuteins, fused proteins, functional derivatives, active fractions or circularly permutated slags are preferred active ingredients in pharmaceutical compositions,

The interferon in the pharmaceutical composition is preferably IFN-β or IEN-a,

In a further preferred embodiment, the pharmaceutical composition comprises a therapeutically effective amount of TNF-α. The pharmaceutical composition of the invention may further comprise one or more CXX inhibitors.

The 2 O · «gyögyászaúfeg elfegaAmő ⁴ 'refers to any carrier competent, which does not interfere with effectiveness of the biological activity of the active ingredient and is not toxic to the host to which it is intended to. For parenteral administration, for example, the active wheal volume can be mounted on a tonne of scintillation dose in a carrier, e.g. in saline, dexfrose, eye albumen or Einger25,

The active ingredients of the pharmaceutical composition for wall deficiency may be administered to the subject in various ways. The routes of administration may be imradenial, transdermal (e.g., slow release formulation), mtramuscular, mtraperitoneal, intravenous, / / ubkutan, oral.

# ** ♦ X9 ·. <9 '* X · »V ·> X *»> Λ> «· <· V

-26MyraNesmialis, Epidural, Re-alcial Topical and Intranasal, Bamayal _{: A} pharmacologically effective route of administration can be used, for example, absorption via epilelial or endothelial tissues, whereby the patient is encoded by the active ingredient D \ 'Sm. vector), which results in the expression and secretion of the active ingredient in Gw. The protein / protein of the present invention may also be administered in conjunction with other biologically active ingredient (s), such as pharmaceutically acceptable surfactants, binders, carriers, diluents and carriers.

For parenteral administration (e.g., intravenous, subcutaneous, intramuscular), the active protein / protein may be formulated as a solution, suspension, emulsion or lyophilized powder with a pharmaceutically acceptable parenteral carrier (eg, saline solution, manna) or chemical stability (pk preservatives and buffers), The formulation is sterilized by commonly used techniques,

The bioavailability of the active protein / proteins of the present invention can also be improved by hoppn gelation methods that a. the folate of the molecule in the human body is well-known. for example, the coupling of a molecule to poly (ethylene glycol) as disclosed in PCT application WO 92/13095.

The memtyiséae effective in pulling off active whites / cows are supported by many factors. IUGG. for example, the type of anagonist, the affinity of the anagonist for 1L-18, the astonishing effect of the antagonists, the residual, eitotoxic activity, the route of administration, the clinical condition of the patient (including the requirement that endogenous 1L inactivity be non-toxic). )

A therapeutically effective amount is an amount which, when administered, inhibits the biological activity of 112-18 by inhibitors 11-18. The dose administered to the subject

- in single or multiple doses - depends on many factors. for example, the formakokinetic properties of the IL-IS inhibitor inhibitor, the mode of administration, the patient's condition and characteristics (sex, age, weight, health, size), the degree of symptoms, the treatments being administered, the frequency of treatment and the desired effect. The common dó »* ♦:» * * # * * ·: <. <

adjustment and treatment of these domains can be performed by one of skill in the art and viral and mww methods for determining inhibition of IL-18 in your child are known to those skilled in the art.

According to the invention, the 1L-IS inhibitor is about 0.0001-109 mg / kg or about 5 0.01-10 kg / kg body weight, or about 0.1-5 mg / kg body weight or about 1-3 mg / kg body weight. body weight or about 1-2 mg / kg body weight of alk. it is so desperate; IL-1A-inbib strain (about 0.1-1000 pg / kg body weight or about 1-100 pg / kg body weight or about 10 -10 .mu.g / kg body weight).

The preferred route of administration according to the invention is subcutaneous administration. Intramuscular administration is also preferred according to the invention. Preferably, the IL-1S can be administered directly to the site of action by lymphatic or intramammary administration. Intraeranal administration is advantageous, particularly in the case of open head injury (bullet injury to the brain).

In other preferred embodiments, the IL-18 inhibitor is administered daily or min1S den on the second day.

The daily dose is usually administered in divided doses or in Tons of sustained release in an amount effective to achieve the desired effect. The second or subsequent administrations may be administered at the same dose, or less or greater than the initial or individual dose administered to the subject. The second or subsequent administrations may be administered prior to or prior to the disease kufakuldx-i;

According to the invention, the IL-18 inhibitor can be administered for prophylactic or therapeutic purposes; before, during or after other therapeutic treatments or agents (e.g., treatment with multiple agents) in a therapeutically effective amount, preferably with interferon and / or TNF and / or other anti-inflammatory agents, e.g.

25: In combination with a C0Xinitrogen and / or anthroxidant. Depending on the brain injury, one for THF

Concomitant administration of TNF analogues is also contemplated (Shohann et al., 1999).

The active ingredients to be administered at the same time as the other therapeutic agents may be administered in the same formulation or other formulation.

* ♦ * «* * * ♦ * *» »** ♦« ♦ * * - ** ♦ «« «*« ψ ·> '** »» ♦ * A »Χ

-28Α. The present invention further provides a method of preparing a pharmaceutical composition comprising admixing an effective amount of lL-1884ohlbltnr and / or interferon and / or a TNF antagonist and / or COXinhibifer with a pharmaceutically acceptable carrier.

The present invention relates to a method of treating CNS sera by administering to a patient a therapeutically effective amount of a horse inhibitor of IL5, if necessary.

All references cited in the description are examples! journal articles, extracts, unpublished or unpublished United States or foreign patent applications, issued United States or foreign patents, or any other references, are hereby incorporated by reference in their entirety, including all particulars, tables, figures and with text. Further, the entire contents of the cited references in the references cited herein are incorporated herein by reference in their entirety.

Reference to known process steps, conventional process steps, known methods, or conventional methods does not imply recognition that any aspect, description, or embodiment of the invention has been disclosed, taught, or disclosed in the art.

The following description of specific embodiments will fully illustrate the overall nature of the invention, so that others may modify and / or adapt the specific embodiments, without undue experimentation, using prior art (e.g., the contents of the references cited in the description? Such adaptations and modifications to the disclosed embodiments may be considered to be identical in meaning and scope to the description and scope of the disclosure. It is to be understood that the terms or terms of the description are intended to serve the purpose of the description and not to be restrictive. It is understood that the terms or expressions in the description will be interpreted by one of ordinary skill in the art in accordance with the teachings and guidance provided herein. yetemben.

Following the description of the invention, the following examples are provided by way of illustration and not by way of limitation.

-2foo · * · *

EXAMPLES

Materials and Methods

Traumamodell

Mice used in the study were 8-16 weeks old. They were 3O-35g mice per month. O5 was bred in a non-pathogenic environment and kept under normal temperature and light conditions, with 4-6 animals per cage, with food and drink as desired. The investigation into Jerusalem! We have been instructed by the Institutional Animal Care Committee of Ifehrew Lníversity (Israel). Experimental CHLt a. with a previously developed weight loss device (Chen et al.,

1906), Briefly, after being exposed to ether, a longitudinal incision was made at the centerline, the skin was peeled off and the skull was exposed. We identified the left frontal frontal area and placed a pointed power cone to the side ~ 1 Mni from the centerline to the central coronal area, the head was fixed, and a weight of 75 g was dropped on the cone with IS cm high, a local injury to the left hemisphere. After the trauma, mice were oxygenated with 100% oxy15 for no more than 2 minutes and then returned to their cages.

To quantitatively assess the iniracramal level of lt-18, mice in the C57BL / 6 (Skin) strain (n ™ 62 in total) were divided into six separate groups: (1) normal colooxo20 porous, i.e., untreated Bo0 mice (n ~ 10), (2). ) an ether anesthetic group where mice are anesthetized with ether. W at the minute, and after 24 hours (n ~ 10) or 7 days (n I), the kidney was slaughtered, (3) a sham operation group, these mice were anesthetized and a longitudinal incision was made on the scalp, followed by 24 hours (n ~ 15} or After 7 days, mice were sacrificed (Group 4i, experimental Cl0 were performed as described above, animals were decapitated under ether anesthesia at 4 hours, (n-7), 24 hours (n 7) and 7 days ( n-7), (5): To evaluate the role of TW in the regulation of IL-18 in the IL-18, the mice were anesthetized with ether, injected intmcerebro-vetricular (10 µl) with 10 µl of sterile, intramolecular Pnepheer saline ( ), 200 ng mouse ·· detn, recombinant TNF half (RAD Systems, Abingdon, UK), and 24 hours later, the animals were killed (n 10), (6) self-injected, and the animals were injected with icv only vehicle (W ui). sterile MS) and then 24 hours (n ~ 6), thus serving as a control group for TNF-injected mice. In all mice, the brain was removed immediately after decapitation, frozen in flowing nitrogen and stored -TOAVoo until assay. Brains from the trauma group were separated into left (ipsilateral) and right (contralateral) lobes to compare IL-W levels in the injured and non-injured hemispheres. Bomogenization of tissues was performed by Mjwk (Kinematiea, Kriens, Svritzerlaud) (1). M) ice cold: extraction buffer in 50 mM Tris (PI 7.2), 159 mM. sodium chloride, 1% Triton-X-Dö (Boeirringer Mannheim, Rotkrenz, Switzcrland) and protease inhibitor cocktail (Boehringer Mannheim). The bomogeuátumot rázaítnk 90 min on ice and then for 15 minutes at & cenin vortexed and spun at 3000 g VA, 4 ⁰ C, the supernatants were divided into small volumes and stored at -70 ° C until assayed. All of the proteins in brain extracts are machineryed

Was determined by Bradfbrd maturation (Bio Rad Laboratories Munich, Oermany) and was found to be the same in all mice tested (<12.1 ± 2.1 mg / ml mean ± mean deviation). specific ELISA according to the manufacturer's instructions (RAD Systems, Abingdon, UK). The sensitivity of the assay is 5 pgrtnl. All concentrations below the detection limit of 5 pg / nrf were labeled with 4.9 pgZnd between different groups of animals for the purpose of comparing intra-uterine IL-18. Samples were diluted in two parallel wells and the final machine center calculated from the mean ÖU of the two samples. OD was determined using a spectrophotometer (Dynatech Laboratories Inc., Ubanilly, VA) at 495 nm extinction wavelength.

SdbBIAkc ^ Bdmbter

Male specimens (h - 40) of the Babie strain of Liebrew University are used for ik-LSBP water bodies A ^? and anesthetic CH1 and experimental CH1 were performed as described above. For treatment, the animals were divided into two groups: Group A (b * * * ♦ * »

Φ «X * * ♦« »♦: ι>

* * φ ** ** «

-31ro & söportx η = horses) were experimental Cffi performed and vehicle-only (PBS) injected for one hour, neurological clearance? Pap was performed (see below) Λ Bc \ oport (exams, group ui, n-ISI) mice were injected with ip 50 pg l -KBP immediately after the neurological score was determined 1 hour after Cl-H. blood brain barrier permeability is 5 to 6 times higher in M 'hours following CHI than previously demonstrated in the same experimental model (Cben et al., 19961 _: IL-18BP can enter the brain under these conditions) Two additional groups of mice were treated with sera A and B (group C: control group, group D: study group), and 48 hours later, the mice were decapitated and the brain was removed and posttraumatic

I0 edema was evaluated as follows.

A neurological severity score system (NSS, Neurologic Severity Score) was developed and validated for the evaluation of post-traumatic neurological damage (Stahel et al., 20G0).

The scoring system evaluates 10 individual clinical tasks of motor function, animation, and physiological behavior such that in case of task failure, one point is counted and: zero point if the task is successful (Table 4). Maximum 10 points: 'NS'S severe neurological disability. means because all tasks failed, ntlg. zero value is for healthy, uninjured mice, measured for 1 h following the trauma

2Ö NSS reflects the initial severity of the injury and shows a strong correlation with clinical outcome (Som-Adaní et al., 2001). Task evaluation was performed by two researchers who did not know the study group (blind study) at 1 hour, 24 hours, 72 hours, and 7 days after the experimental CBI, the ANS.S measured at 1 hour by NSS and any other , the difference in NSS measured at a later time point is a parameter indicative of the degree of spon25 recovery after brain injury, as previously described (Cben et al.,

1996).

'3 · Ύ

444 4 * * «* * ·« «

Table 4:

Heurological Nervous System (NSS) in mice with head injury

---------------------'---------------------- Task .eiras

Fonts Success / Flipping 1

Exit circle: | Mom-Zheiniparesis ií | That ability? 1 is a 30 cm The opposite is ol and initiative 3 minutes from lap » able inside the tendon sky. that contact 0/1 :: dali top and / vi rod rgy lower water very good 0/1 09 , ψνί-ί Á-.l A Ύ VÍ.A? 1 1 ability g to walk straight í J Fright! reflex | Veleszületettre fi ex, the mouse h angos íap> , .ra jumps. 0/1 í Search behavior i j The environment is Irish Inti inquires; behavior st indicator phys iölogiai 0/1 | Rod Balancing: 1 7 mm wide; i breaks through S rod to the bottom damn straight scarf) fe 10 rmfo Image of Ozas rdpercen population 0/1 i 1 With a round stick | 5 mm in diameter. circular pile; tendon at the bottom ? 10 sec 0/1 1

teno cg \ self-weight balancing through o / s and walking on rod in cm; 30 c., 3 sts wide on rod. | ability to walk on em rod j Same task difficult with 2 cm wide rod 1 cffl-cs rod walking 1 gsauez on task 1 cm Rod with uebe / nve Maximum point

Evaluation of bedding

The magnitude of eerebral edema in the hemispheres was evaluated by measuring the water content of the tissues as described previously (Chen et al., 1996). Short

-33mice were described above with the Serbian etiquette 48 hours after the trauma, which is its. corresponds to the time at which edema is still significant in this modelbends / crben IChcu> .s rusm. 19 ^ 6; After feA '! E / kwugwü and a / aguet / se * have been removed, the area adjacent to the site of injury and the

2b mg up to one! participate in our little self. The right (uninjured) bowls were used as an internal control. The tissue sections were weighed and dried at 95 ° C for 24 hours. Dried safes \ U) at 'emermx the volume of egv ϊν, .ωΙΑο ⁴ · is hoseske, ok we shopped it% H? O - [wet weight: - dry weight] x 100j /' wet weight: Brain injured

W We studied patients with physical, severe CHi who were admitted to the accident department of the Ihúversity Oospital in Zurich (mean age ± mean difference: 37 ± shoot years, age range 24-5 years, 9 men and I women). All patients had a Glasgow Gore Scale (GCS) of <8 after cardiopulmonary resuscitation (Tleasale and Jennett, 1974). After CT examination, all patients received an intraventricular catheter for therapeutic CSF drainage when intracranial pressure (1CP) was greater than 15 mmHg. You haven't treated any patients with steroids. Patients with multiple injuries who have had chest, abdominal, pelvic, spinal cord injuries or interventions due to fractures. It was also necessary, we were excluded from the study. Individual consequences were estimated on the basis of the Glasgow Öuteome Seale (GÖS, Glasgow Consequence Scale) (Jennek and Bond, 1975), at CSF'node. and serum collection was approved by the Ethics Board Committee of the University of Zurich, Hospltal,

The CSFs of patients with CHI (nMO) were collected daily at a specific time on the corresponding serum cold. A. Control-CSF was obtained from patients with diagnostic spinal cord drainage (n ~ $). These patients showed no evidence of CNS inflammatory disease based on normal CSF values for protein, glucose and cell counts (data not shown). In GHI-esoportan, collecting onutut 10 days of trauma kmetou '\' gertmk b tesw not for \ erui \ nl m- who fom, feun wol cakes ek pl uhun

-34 * á »·» *** <

* ♦ ♦ X * ♦ * »* * x»

X X-X tr ♦ «* jfr

M ♦ ♦ «w>« fe * »in cases where 1CP remained at normal value (<15 Bgínrry for more than 24 hours. A total of 1.000 compatible CSF and trauma patients in this study and All samples were centrifuged immediately after collection, aliquoted into small volumes, and stored at -50 ° C until assay. A quantitative assay for CSF and serum II-18 was performed on human lt-I8 specific EL. ISA was performed with a commercially available reagent kit (R&D Systems, Abingdon, IL) .The sensitivity of BUSA to mouse assays was 5 pg / ml, and the final IL-machine centrifuge was calculated as the mean of 01 duplicate samples at 405 mn extinction wavelength. To compare the 11, -18 CSF stains of CH1 patients and controls, all concentrations below the 5 pg / ml detection limit were labeled with 4.9 pg / ml males.

Statistical analysis of the data was performed using commercially available software (SPSS 9.0, Windows ™), Non-standard distribution data (eg, NSS and ANSS neuro'15 logical results) were used with non-parametric Mann-Whimy ~ U tests, Unpaired Stndent- Half of the mice were used to compare the inferior IL-18 concentrations of different mouse groups and to examine the differences in brain water content from IL-18BP treated and vehicle treated mice. Comparison of human IL-18 levels as well as daily CSF in patients with CP11 and

2 <0> corresponding serum samples, as well as the Comparative and Contrho-CSF-ck comparisons with the ANÖVÁ general linear model suitable for repeated measurements, A <0.05 values were considered statistically significant.

EXAMPLE 1: IR-18 IL-18 Figure 1 shows that IL-18 ELISA was able to detect a plethora of untreated (normal) control mice (brain 10) with an average level of 27.7 ± 1, 7 ng / ml tt SLMj. In the experimental groups: the same with ether anesthesia alone or with anesthesia with sham surgery (ie ether anesthesia and longitudinal head circumcision) significantly

IS

Φ fc X fc ♦ X # *

Φ * resulted in a higher intraeranal IT-ld lesion: 48.9 ± 1.1 ng / ml (ether: group, η ~ §) and 54.3 ± 2.7 ng / ml (pseudoperative group, n ^:::: 13} (p <δ, kills versus normal mice, unpaired Stndent test, Figure 1) .The difference between animals in the ether group and animals in the sham operation group is not statistically significant (p = 0.16).

In the subject group (n ^:::: 21), Cili triggering increased: IL-18 levels in both & injured and contralateral passages 4 hours after trauma (60.6 ± 3.3 vs. 59.8 ± 5, 0 ng / ml} to 24 hours (56.9 ± 2.1 to 56.3 ± 3.7 ng / ml), however, these levels were not significantly higher with respect to the ethereal or transoperative groups (p> 0, : 05).

In contrast, at day 7 after CH1, concentrations in the injured hemisphere were significantly higher than those of ether-anesthetized or sham-operated animals (? 6 ± 5.1 µg / ml at 42.2 ± 0.8 ng). / ml and 45.2 ± 0.5 ng / ml, p <001). whereas in the opposite hemisphere, IL-18 / tendon did not significantly increase above the values in these two control groups (63.2 ± 6.0 ng / ml, ρ ^::: {sight}).

TNE ,. in this transmammal model, an additional group of Bö mice (n ~ 10) was injected t.c.v. to assess the role of an important mediator of inflammation (Shobanh et al., 1999) in the regulation of intra-uterine IL-15S. The animals were sacrificed 24 hours later with 20 µg / ml of µ μΐ sterile BBS in murine recombinant THF. Figure 1 shows that self-injection of vehicle alone (n = 6) resulted in a significant up-regulation of intracerebral 1L-18.

Within 24 hours, compared to untreated normal B0 mice (53.6: fc 3.9 ng / ml versus 27.7 2 1.7 ng / ml, p <0.001).

Injection of TNF resulted in a significant reduction in IL-18-smm within 24 hours (22.1 ± 6.9 ng / ml, n = 10) of the mtraleramic compared to the 24 hour value of the injected conirol group (? Lo ± 3.9 ng / ml). ml, p <0.901). The levels of l.L-1825 in the TNF group are not yet. .hops, were also lower than normal mice (2.7.7 ± 1.7 ng / ml), although in this case the difference was not statistically significant (p ~ 9.45).

^ .ItdlBBfedé

In order to investigate the theory that inhibition of IL-1S promotes healing of the brain, healing was performed at different times after a single injection of 11.M & 13F. Previously shown to be neurological severity! point system (NSS); one hour after the trauma most accurately reflects the size of the trauma and the amount of tissue damaged, as seen on MRI histology.

Mice were divided into different treatment groups at t ~ 1 hour for baseline NS-S to produce comparable levels of traumatic mice. As shown in Figure 2 [HSS in IL-1 hBE group (squares) versus kmhroll group (circle)], both groups had similar initial NSS (1 hour) values (7.9 ± 6.3023 (control) and 7 , 44 ± 6.3027 (IL-1 SBP) j, and indicates similar severity.

Subsequent evaluation of the NSS (1-7 days) showed that animals treated with IL-1SBE with intraperitoneal (i, p,) significantly less

IS damage, as seen in the NSS values, and significant difference 7 days after trauma (ρ 0.645).

The healing rate was calculated as ANS8 (u ^::: NSS (I h) - NSS (t). Higher values of ANSS indicate faster healing, zero or negative values indicate no or worsening healing. The difference between the mean of the ANSS values at two times, 24 hours and 7 days, is significant.

Another experiment was conducted to investigate whether starting IL-lkBP was effective 3 days after injury. A. The timing of the onset is important and so far they are. therapies that have been started within a few hours have proven effective. Because we found that IL-1S · rises 7 days after trauma, and lt-18BP ~ ve! Treatment 1 hour after the trauma results in the greatest effect even on day 7, we decided to give the mice 3 days a. treat the injury afterwards. The Authentication * *

-37 · another group was treated with IL-8BP for 1 hour and 3 days for woody. Control group A was treated with solvent (vehicle) only.

The results of the experiment are illustrated in Figure 4, which shows that a. A single treatment on day 3 is as effective as 1 hour after OII and then on day 3.

day repeated treatment.

The experiment also demonstrates that at a given time, IL-1. & BP, either at 1 hour or up to 3 days after a closed head injury, exerts an extremely favorable fish model in its mouse model of recovery from transient head injury Macasabb.lQ $ ό mtek ag> sertó atanthuntán OCN

Ten daily CSFs from ten severely affected CBIA patients. and IL-18 levels for tenth day after trauma, and demographic and clinical data of patients are shown in Table 5.

Ádábfezat

Serious αίems suffering bеgem demography ^

Patient number Disease (Years) feem GCS ^b GOS ^C 1.1 ..- 18 in CSF ^d (pgórd) l.L-18 in serum) average [Provincial * average [íartománvf 1 48 / nt 3 1 283 [78-966] 57.5 112-661 2 31.ón 7 4 228.4 [22-745] 19.7 [4.9 to 108] 3 26 / tn 5 3 208.5 [20-392Ί 37.2 [14-1631 4 57 / m 5 4 72.6 [30-2861 48.9 [14-1041 5 24 / m 4 5 32.6 [4.9 to 155] 17 B ^ 8] 6 3 sins 8 I 49.4 [10-290] 16.7 [12-67] 7 38 / f 3: 4 17.9 (II-lOOj 13 [4.9 to 46] 8 35 / m 3 3 69.8 [4.9 to 329] 19.8 (7-771 9 3? / M 3 4 37 [23-75] 57.3 [25-98] 10 4I / m 7 5 4.9 [4.9 to 169] 26.2 [15-381 Control CSF (n- ^: 5) 4.9 [4.9 to 7.8]

-38 ^Λ € Β1, dosed head iniury ^ GCS, Glasgow Gore Seore · '(Teasdale and Be,

1974) ^C GÖS, Glasgow Ouíeome Sörre, 3 months 5 after injury, 5 ”asymptomatic, 4 ~ moderate dysfunction, 3 ^:::: severe dysfunction, 2 ^:::: persistent vegetative state, I ~ death (Jehnett) and Blond, 1975) \ SG eerebrospinal fluid * IL-18 levels below 5 pg of detection are designated as 4.9 g / gal.

Shoot

As shown in Table 5, intrathecal IL-1S staining was significantly elevated in 9-Ί0 (.illm) patients compared to control CSF taken from 5th patient without trauma or inflammatory neurological disease (p <δ. measurements (ANOVA). Only one patient (lead) had no significant increase in IL-18 CSF in control

Relative to CSF (p = 0.31). The average AA levels and the individual ranges of IL18 sximes measured in CSF and serum are shown in Table 5.

It is noticeable that CSF with maximal IL-18 machinery (900 ng / ml) is up to 200 times higher in patients with head injury than in conflates. Intraeepheneal IL-18 FLIS A was detectable in 9.0% of the CSF samples in the feaumaoseporthau, while 70% of the control-CSF samples had detectable mRNA. IL-1 S-smi (i.e.:>

4.9 hp mf Λ per mole of Glll-seven cosid m m average IL- ^? 8-koncenuac ok sumumbLawan higher; were in the CSF. mii in serum (p <He, Ö5, repeated measures ANOVA). however, in beta patients (# 9, shoot), mean serum 1L-18 values are nern. were higher than the concentrations in the initial CSF as shown in Table 5.

As a result, IL-18 levels were significantly elevated in the eerebrospinal fluid of patients with head injury due to urticaria. Addition of IL-1SBP can reduce these elevated levels and thus have a beneficial effect on healing from a closed head injury, as described above in Example 2.

39 * ♦ * í

M »«} $ ♦ * 9 * X # V $ «** 9« «9 X-9 #

References ü af ot <4 to * m 215, <$$ hd s F fe <Htedfec _f 2S>

0SB <R5x4 ^ S4),

3, Chsn, V- B <Constendnj V, Trombovteh M, and E, Shdmú. 1

ώ. <^ $ sss «3 $> TV,.

> í * &&

fe feaood honé fetey fe r *% j6Ö

4, Cor <C.8., H, Y. Cslíngnsnso Y. Kim, H. fem ₍ Y. δηη, £, Glbson, nmá Ϊ.Η. doh. 1899 «Calteras fe otemoytes aad mternpfe axproan fetefefecfe-í8« Mole, 3rofe

5. Cood, B. «d> W. dohog, 0, Tinfe 4. H, Són, and TH doh, 1997, Inotedion of r fe ihn sdronoi mfeex 4. Bife Chem, 272: 2038ne, A, C, M, 0th Hell, S J. RodnneC and G.C. UsheshL 1938.

the. Efewtws 4 fe fe, teudteo Afeds Re®, 12, SS7-3S5,1984.

'? .. B§3eneM 4 A, Hevstew®, M _s Mw <ö M, Zondi, £ feg fe ffe.

jí «V <.5: ΐ

8. Bited M4L, teteni, RM, Fektfearfe, Ufeg-Fox, A, Ctertsa, F, Bijk

1894, Laecfe 344,112S-1127, fed K _s O, tetel .. T, Becs izutv, ode 8,% w. 1999, h feterfeme-y fe feOanwYory CHS dteoaa 10. Jancfc.r, S, _s and G, Stete 1008. Wfeetei

IS, and inteteootetete swefeag onows mRHA fe t nncophstennytebte felbe Lmafe afe., Λ Hfeeölfefeeaöl, 91: 83-9 11 Lenem 1975 Mer 1; 1 <790S}; 4S £ M. 4mOO 8, »h M.

13. Male: SH, Befetfeefe M judetlkev t, Farfefe S <

ft <18 bofeg protein te white-18, Froo Rfel Acad Sd ü É 2®W:

83: 1104-8 M, well.

XR x «*>: 4

-46 »* <. * 4 *

4 4

X- ν:> «* ·>

9 4 »* * tfy« V

Kosomasm X, P, F, SfeM, Ρ .Μ ΙΑ.

IsfeMb.

18. Kbight IM Tddh K Le A SfegM S, SbWy O _t

MA Weesk A öddona P, et <Confemfeín séd MsÉ öbarssfe-udon ef a monse-bunw ehímede- endXHF aotModyMfe rtoM 1M Nov 38; 161443-53

16, Mfeiszewnfe ,, C. R., X A. Safe, X Vaodeo Boa,. S, Wsegh, S, X .. 0 <w «A SMsk, Μ, R. BecAmanfe dodo K. Hl Grabstefe, 18541. CytcKse rooapfers and B oaS IR fehÁA <« 1 «and ÍMtak I.

Mmssd Ö eeí atMte b vfeo, A Rím 17, IfeM »A, Ts O8AM8, K. Mm P. X

Tawhófe, K,

sMMMM human 7 «Μ synerAsm wffe tnfedauWIS far fe AAterd M.1847A1 Mm 8814-2880.

M.G,> PM tanfenguu V, Hsns, P „3 fen, P- GfecMr, Ö> Tmnfe, and T,. Kossmsnm W. P cRMfeas feíMMg brafe bjwy. bwtei and defeíer «χ ·.

White, W) C

H, Wedd PL Nagfea K, (1O), Immynfey 16 _t 127-7

21, Ökumufe H, Hagafo K, MmaM X Ta? Tefego © K, ekor T, Fos

22, PorM, P, Gafea, X £,. fened, T P, Chem, 247 WW ¥,

3 (10): 3388-72 Dower, B K, ®d SMs, d B, (1 §§S), A 8M

a, PeatwíW R, MethsM b Bs ^ hxjfegy, 183.83-88.1980 h. Marson W R and öpman 0 A Pnoo Xat Aeed 88 OB

23, Rlort ki, and UK Hanísoh, 1Ό, fe fefedatAkMe. The New

'* »* $ ♦ * *« *> fc

AZ> fcfc *.> «Fc * _A ^r X <fc fc» ** ** * f> ¥ · **

-41: ♦ ** »

'sipba in örafe ir ^ nty, Cytekhó Gmwfe Bas & r Bsw tö; 11 2 «30 27, Shoh & rb, E; BsíOVannal Homwiíz M; Koten R (1997); In Comb, 8

IX 16W-1W.

£ §a »í'S. irfWífcSfcÖíí ^ '£> $ “fcfit'Tjií Sfcíiíi vt, .Urtfsr« _s anf'-Kossnwn MU J

Ft

GT

K, V, AM & fC $ M, Tongoe K _s Tardmoto T _s Fteuda S, iketfe M, Oxmura mffteb 1333 3 w 1í1 $ S (11) s <274-9 feri R.Ö., A3X Üblhsss, MÖ, RMl, S. , RsteH ^ Wöáfflk íUWM 2Oöa Detecbm of the btetfelAWS Mrbly in mt ohrain by ier ,, Mj _v TM Carfcs, FA. Kod-wtek, SR, WboMwMd, ód Bell,;, ST. GeKoedy, G, W, kladnn, and PD, Adebcr ', 2000. feíerfeufe · in csmb'mpinsi ifelö of chiidmn wtth ssavare head Injury. CM. 23: 929-34 nfea X Takeá®, K _f Tanaha, T ₍ ÖóteM, iOX 3. Immurfo. Wi 340 (93407,

Claims (10)

Use of a 1,1L-1β inhibitor, which is a 5L-t3-α fatty protein (OL-18 O), for the manufacture of a medicament for the treatment of typhoidal brain injury, administered as a single dose of Ib-1S BP 3 days after injury. . The use according to claim 1, wherein the trsomatic brain injury is a closed head injury. The use according to claim 1 or 2, wherein the IL-1b BP is glycosylated at one or more sites. 4. The use according to any one of claims 1 to 5, wherein the O, -18 OP is fused to all or part of the immunoglobulin (µg). The use of claims 1-4, which is a Serbian application, wherein: the IL-18 OP contains at least one moiety linked to one or more Smcoalic groups on one or more eclectic residues of the amino acids. The use according to claim 5, wherein the molar portion is polyethylene glycol (PBG). Use according to any one of the preceding claims, further comprising a medicament. The use of claim 7, wherein the interferon is interferon-a or 'interferon-fk SHE. Use according to any one of the preceding claims, wherein the medicament further comprises tumor necrosis factor (TNR). The use according to claim 9, wherein the INF is an INF. Use according to any one of the preceding claims, wherein the medicament also contains an anti-glaucoma active ingredient, 12. All. The use according to claim 1, wherein the gynophilic antiinflammatory agent is COX-Inhibitor, preferably COXd-Inhibitor. 1.3. The use according to any one of the preceding claims, wherein the medicament further comprises: The use according to any one of the preceding claims, wherein 0.1 to 18 BP is about 1, W 1 to 8 mg / kg. or about 8.84-10 mg / kg. or in an amount of about 0.1-5 mg / kg or about 1-3 tttgdifcg. The use according to any one of the preceding claims, wherein the 4L-18 OP is administered sncubically.